Western blot analysis of C-Kit [pY568/pY570] was performed by loading 20 ug of A549 (lane1) and Serum Starved A549 (lane2) cell lysate using NuPAGE® 4-12 % Bis-Tris gel (NP0322BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. C-Kit [pY568/pY570] was detected at ~ 170 kDa using C-Kit [pY568/pY570] Rabbit Polyclonal Antibody (44490G) at 1:1000 in 5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Rabbit IgG - HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human c-Kit that contains tyrosines 568 and 570. The sequence is conserved in mouse, rat, chicken, cow and dog.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Western Blot (WB)||1:500-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
c-Kit, also known as CD117 and stem cell factor receptor, is a 145 kDa transmembrane tyrosine kinase encoded by the c-Kit proto-oncogene. c-Kit acts to regulate a variety of biological responses including cell proliferation, apoptosis, chemotaxis and adhesion. Ligand binding to the extracellular domain leads to autophosphorylation on several tyrosine residues within the cytoplasmic domain, and activation. c-Kit mutations correlate with tumor growth and progression in a variety of cancers including mast cell disease, gastrointestinal stromal tumor, acute myeloid leukemia, Ewing sarcoma, and lung cancer. Phosphorylation at tyrosine 703 of c-Kit allows binding of Grb2 and activation of the Ras-Raf-ERK1 and 2 signaling pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Increased ERK and JNK activities correlate with disease activity in patients with systemic lupus erythematosus.
44-490G was used in western blot to discuss the contribution of the p21ras/MAP kinase pathway to systemic lupus erythematosus
|Molad Y,Amit-Vasina M,Bloch O,Yona E,Rapoport MJ||Annals of the rheumatic diseases (69:175)||2010|
|Mouse||Not Cited||Lyn contributes to regulation of multiple Kit-dependent signaling pathways in murine bone marrow mast cells.||Shivakrupa R,Linnekin D||Cellular signalling (17:103)||2005|